Abstract:To clone and express the rhoptry protein 18(ROP18) gene of RH strain of Toxoplasma gondii, the genomic DNA, extracted from tachyzoites of RH strain of T. gondii, was amplified by PCR with a pair of specific primers, which was designed according to the encoding sequence of ROP18 gene. The PCR product about 1 665 bp was cloned into prokaryotic expression vector pET-28a(+) or pET-32a(+) with restriction enzymes BamHⅠ and HindⅢ. The recombinant vector pET28a-ROP18 or pET32a-ROP18 was then transformed into E. coli BL21(DE3) and induced with IPTG for expression. SDS-PAGE and western blotting showed that the expression product was a non-fusion protein about 60 kD with pET28a-ROP18 and a fusion protein about 83 kD with pET32a-ROP18. The antigenicity of ROP18 was detected by western blot with primary antibody of prepared mice antiserum against T. gondii. With the immunological effect of this expressed recombinant protein, the present study might provide the foundation for the further study of ROP18.
王素凡,齐辰,陈滨,吴焜,陈晓光. 弓形虫棒状体蛋白18的原核表达及免疫分析[J]. 中国人兽共患病学报, 2013, 29(3): 211-215.
WANG Su-fan,QI Chen,CHEN Bin,WU Kun,CHEN Xiao-guang. Prokaryotic expression and antigenicity analysis on rhoptryprotein 18 of Toxoplasma gondii. Chinese Journal of Zoonoses, 2013, 29(3): 211-215.
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